Diagnostic Laboratory Guide

Tennessee Department of Agriculture Regulatory Services
The Kord Animal Health Diagnostic Laboratory
KORD ANIMAL HEALTH DIAGNOSTIC LABORATORY

TENNESSEE DEPARTMENT OF AGRICULTURE, DIVISION OF REGULATORY SERVICES
http://www.tn.gov/agriculture/regulatory/kord.shtml
U.S. POSTAL ADDRESS OVERNIGHT DELIVERY ADDRESS
Kord Animal Health Laboratory Kord Animal Health Laboratory

P.O. Box 40627 Ellington Agricultural Center
Melrose Station 436 Hogan Road
Nashville, Tennessee 37204-0627 Nashville, Tennessee 37220
TELEPHONE NUMBERS
Information 615.837.5125
Brucellosis Results 615.837.5120
Tissue Receiving 615.837.5410
Serology 615.837.5128
Immunology 615.837.5221
Virology 615.837.5268
Bacteriology 615.837.5427
EAC Security 615.533.9945
Billing 615.837.5410
Blood/Serum boxes 615.837.5231
FAX 615.837.5250
INTRODUCTION
The Kord Animal Health Diagnostic Laboratory (KAHDL) is the State of Tennessee’s veterinary diagnostic laboratory
and is funded by legislative appropriation and fee revenue. It operates within the Tennessee Department of
Agriculture, Division of Regulatory Services to offer no-cost diagnostic services for food animal producers and
modest or discounted fee services for companion animals.
The mission of KAHDL is to provide accurate and timely diagnostic services to Tennessee veterinarians. KAHDL is
strictly a diagnostic service that works in concert with local veterinarians, who are best able to evaluate the client
or producer’s problems and recommend appropriate actions/treatment. It is strongly recommended that case
material be submitted by or at the direction of a practicing veterinarian (although owners may submit cases
directly if necessary). Results of examinations, analyses, and tests will be returned to the veterinarian to formulate
a diagnosis and recommend action to clients. If results indicate the presence of a reportable disease, they must
also be reported to the State Veterinarian. If at any time there is a question regarding a test or procedure, please
call prior to submission to be assured that the service is available and determine specific submission requirements.
Consultations regarding disease treatment are not provided by KAHDL.
We welcome suggestions and comments; we also invite veterinarians and veterinary paraprofessionals to visit the
laboratory in order to meet our team and observe methods used in various diagnostic procedures. These visits
help us understand your problems related to laboratory use and help you in preparing submissions so that optimal
diagnostic information is obtained.
KAHDL Lab Use Guide, August, 2013 Page 1 of 28

TABLE OF CONTENTS
Hours ……. ….………………………………………………………………..………………….. 2

Fees ……………………………….……………………………………………………………….. 3
Shipping Requirements …..………………………………………………………………. 4
BVD Ear Notch Sample Collection/Submission Requirements ……….... 5
Equine Infectious Anemia or Coggins testing …………………………………… 6
Master List of Subcontractors ……………………………….…………………….. 7
Abortions ………………………………………………………………………………………… 8
Bacteriology & mycology submissions.……………………………..…………….. 8
Microbiology users guide …………………………………………………………………. 9
Trichomonas & Campylobacter testing ……………………………………………. 10
Necropsy submissions ……..………………………………..…………………….…...... 11
Histopathology ………………………………………………………………………………… 11
Cytology ....……………………………………………………………………………………… 12
Parasitology…….………………………………………………..……………………………... 13
Serology…….…………………………………………………………………………………….. 14
Available serologic tests & specimen requirements, avian..………. …….. 14
Available serologic tests & specimen requirements, canine..……….….. 15
Available serologic tests & specimen requirements, equine.……. …….. 16
Available serologic tests & specimen requirements, feline.………. …….. 17
Available serologic tests & specimen requirements, porcine….... …….. 18
Available serologic tests & specimen requirements, ruminant...………. 19
Toxicology Tests Available and Samples Required……. ……………………… 20
Virology, tests by species ……………………………………………………………….. 23
Sources of Help in Tennessee ………………………………………………………. 23
Reportable Diseases in Tennessee ……………………………………………… 24
Submission form, Surgical Biopsy ……………………………………………… 25
Submission form, Fresh and Fixed tissues …………………………………….. 26
Submission form, Necropsy ………………………………………………….…… 27
Owner’ Release form, Necropsy ………………………………………………. …….. 28
LABORATORY HOURS
• 8:00 AM to 4:30 PM, Central Time.

• Observed Holidays:
o New Years
o Martin Luther King’s birthday
o Presidents’ Day
o Good Friday
o Memorial Day
o Independence Day
o Labor Day
o Veteran’s Day
o Thanksgiving and the following Friday
o Christmas
FEES (SUBJECT TO CHANGE, EFFECTIVE JANUARY 1, 2012)

• Fees for the Kord lab are determined by state statutes TCA 4-3-203 & 43-1-703
• Livestock testing (used for food & fiber) No charge for Tennessee residents
• Equine testing:
o Equine Infectious Anemia (Coggins) $7/Test for Tennessee residents
o Other (non-Coggins) equine tests No charge for Tennessee residents
• Companion Animals Fees schedule below for Tennessee residents
DETERMINATION OF FEES CHARGED PER SAMPLE BASED ON TESTING REQUESTED/REQUIRED
PATHOLOGY
• Histopathology/Biopsy $35.00 per animal
• Necropsy (no toxicology) $70.00 per animal, limb, organ, or head
• Necropsy (with toxicology) $105.00
• Necropsy (with spinal cord) $95.00
• Cytology $30.00 per tissue (per problem site)
• Slide preparation/Cytology $15.00 per specimen
• Decalcification $10.00 per tissue
• Special stains $2.00 per stain
• Slide preparation/HE $8.00 per slide
• Slide preparation/IHC, charged $10.00 per block
• Submitter special stain $10.00 per slide (requested by submitter)
TOXICOLOGY
• First test $35.00
• Each additional test $15.00
VIROLOGY/IMMUNOLOGY
• Serum/FA, etc. $15.00 each
• Lyme Disease (Borrelia) $18.00
BACTERIOLOGY/PARASITOLOGY
• Aerobic culture $15.00 per swab, tissue, site, or sample
• Antibiotic sensitivity $8.00 per isolate
• Fungal culture $15.00
• Anaerobic culture $15.00
• Stain, gram, acid fast $5.00
• Ectoparasite Identification $15.00 per test
MISCELLANEOUS CHARGES
• Save remains or save ashes $50.00
• Disposal (animal or parts) $25.00
• Leaky package/biohazard handling $15.00
• Shipping and Handling $20.00
• Packaging fee $4.00
• Accessioning fee $2.00
• Faxing by special request $2.00
• Sample supplies Cost plus 10%
• Past-due accounts 5.0% per month

GUIDELINES FOR SAMPLE SUBMISSION
• All submissions must be accompanied by an appropriate submission form which must be filled out completely.
• Live animals cannot be brought to the laboratory for collection of specimens or for euthanasia (except fish).
• Reporting Results
o An E-mail, telephone or fax report will be given on any significant laboratory findings as they become available
or if the result requires explanation by a veterinary diagnostician. Please make sure the submission includes
both an Email address and fax number to avoid delays (there may be a fee for faxed results).
o A written report will be sent to the referring veterinarian upon completion of the case. The preferred
distribution method is by Email. If a fax or USPS mailed copy is required, additional charges may apply.
• Copies of the owner’s written report are sent to the owner if the owner delivered the tissues and/or animal directly to
the laboratory or the submission is from a farm animal species and the owner’s complete address is present on the
submission form. E-mailed results are preferred, copies sent by USPS will significantly delay results.
• Please report any changes of Email, address, phone number or clinic association to the laboratory at (615) 837-5125.
• Veterinarians utilizing laboratory services may choose to send payment with submitted specimens or establish a charge
account with the laboratory. Accumulated charges will be billed to the clinic on a monthly basis. Accounts 90 days
overdue are subject to both a withholding of fee-based services until payment is received. Credit will also be
discontinued requiring payment accompany all samples.
• Submitting samples, specimens, or animals to KAHDL implies agreement to policies of the State of Tennessee and
KAHDL and transfers ownership of samples and property to KAHDL upon receipt unless otherwise determine by written
agreement prior to submission.
• Safety of laboratory personnel is paramount. Submissions with attached needles, leaking formalin or other fluids are
subject to being discarded or may have a biohazard fee levied. Absolutely no human samples are to be submitted. For
other primates, only formalin-fixed samples will be accepted (absolutely no fresh tissue or fluid samples from primates).
• Animals that are submitted with the concern or suspicion of being shot must submit radiographs (X-rays) of the body
part in question in two orthogonal views if retrieval of a projectile or confirmation is expected. For companion animals,
additional charges may apply if additional documentation or testing is required.
• Bone lesions or malformations are best diagnosed by radiographs. The laboratory does not have radiographic
capabilities and has limited capacity to examine bone lesions.
• Formalin-fixed tissues are required for histopathology examination. We do not accept fresh tissue for histopathology.
• Brain for rabies testing is best send to the Health Department’s rabies testing lab. Specimens sent to KAHDL will incur a
necropsy fee to help offset the costs of sample procurement, processing, and shipping to the Health Department.
SAMPLE SUBMISSION BEST PRACTICES
Serum - Collect blood aseptically into a sterile dry tube, refrigerate immediately, allow to clot, centrifuge, and transfer
serum aseptically into a second tube. Serum must be fresh, clear, unhemolyzed, and uncontaminated. Using red top
®
Vacutainers , B-D, or other non-EDTA/heparin sterile commercially available tubes are ideal. Do not let whole blood freeze
or expose to direct sunlight or high temperature before decanting serum.
• Label each tube (not stopper) with tube number and vet code. Be careful that writing will be legible upon arrival (use
an indelible marker). Identify specimen in a way that will avoid confusion when results are reported. Avoid using
animal names. Keep your own log of sample numbers to avoid duplication and confusion. Be sure to number samples
consecutively and maintain a master list. Always include a copy of this master list with the submitted sample. Put small
tubes in a box or tape to large cardboard so they will not be lost in the packing materials.
• Always call if sending more than 25 samples so that the laboratory can prepare for extra cases.
• Submit at least 1.0 ml of serum for each test requested. Refrigerate the serum until shipment.
Fresh tissues should be collected aseptically and placed individually in well-labeled Whirl-Pak® bags and refrigerate
immediately. Refrigerated samples should be shipped with sufficient cold packs and padding. Ship samples in a leak proof
insulated container or in sturdy mailing carton which complies with postal or commercial carrier specifications. Submit
samples early in the week to avoid holding over the weekend by the carrier. FedEx® or UPS® are preferred. Shipping by
USPS may result in significant delays.

Complete and legible form(s), (including a complete history), must accompany all submissions. All regulatory charts must
include the submitting veterinarian’s signature, vet code, and animal identification.
KAHDL is proficiency tested and certified by the National Veterinary Services Laboratory (NVSL) to perform:
• Anaplasmosis enzyme-linked immunosorbent assay (ELISA)
• Agarose gel immunodiffusion (AGID) for bluetongue, bovine leukemia virus, and equine infectious anemia
• Mycobacterium avium paratuberculosis (Johne’s) ELISA
• Johne’s, Avian influenza, and Exotic Newcastle’s Disease polymerase chain reaction (PCR)
• Leptospira microagglutination test
• Pseudorabies serologic screening by gB ELISA
• Classical swine fever PCR
• Foot-and-mouth disease PCR
Sampling & submission protocol for testing ear notch specimens for BVD:
• Materials and equipment needed:
o Ear notching tool that yields a 1 cm x 1 cm notch. Recommend purchasing two or more tools
 Nasco: 1-800-558-9595. Order ID medium C0024N
 Dairy Health USA: 1-800-276-7933. Order ID medium 440-31
 Stone Livestock ID & Show Equipment: 1-816-231-4020. Order ID medium 7125
o Red Top blood tube: Vacutainer® 7 ml draw 16mm x 75mm collection tube with no additives is preferred.
o Shipping container that holds individual tubes in slots.
o Submission Form from the Kord Animal Health Laboratory (615-837-5125) or from the end of this guide (at
http://www.tn.gov/agriculture/regulatory/kord.shtml)
o Disinfectant for rinsing notching tool: 10% bleach (100 ml (3 oz.) bleach in 900 ml (27 oz.) water).
o Clean rinse water: 3-5 gallon bucket. Change bucket water every 20-30 notches.
o Disposable gloves and clean coveralls.
• Collection & submission procedure
o Label Red Top collection tubes with consecutive numbers (#1 through number in submission) and keep a list of the
corresponding animal ID or tattoo numbers. This method of labeling significantly reduces the turnaround time in
the laboratory and gets the results to you faster.
o Dip notching tool in disinfectant, then ALWAYS rinse away disinfectant with copious quantities of clean water.
CAUTION: Residual disinfectant on the notching tool will yield false negative results, therefore thorough rinsing
with clean water is required!
o Collect an ear notch approximately 1 cm x 1 cm (3/8 in. x 3/8 in.) from a clean portion of the ear. Only fresh ear
notch samples are acceptable for the antigen-capture ELISA test. The ear should not be cleaned with anything but
water. Disinfectant of any kind (e.g. Novasan) can interfere with the test.
o Place the fresh ear notch into a sterile, clean Vacutainer® collection tube (no formalin, saline, bleach, other liquid,
detergent residue, or separator gel). Do not use Whirl-Pac® or any other type of plastic bags. CAUTION: Collected
ear notch must be free of contaminating dirt, feces, tattoo ink or BVD vaccine. Do not vaccinate or tattoo at the
same time samples are taken.
o Send samples to the laboratory with sufficient ice packs to chill the specimen during shipment. Saline or other
media must NOT be added to the tube containing the ear notch. Likewise, please do not use any fixative (e.g.
Formalin).
o Notify the laboratory in advance prior to shipment regarding the number of samples and when they will be sent.
This allows us to have adequate supplies for testing and to arrange staffing to do the testing.
Equine Infectious Anemia (EIA or Coggins) serology requires at least two (2) ml of clear, fresh, non-hemolyzed serum and
takes at least 24 hours to complete when the laboratory is fully staffed and sample numbers are not excessive. Retesting
may be required. Each tube of serum submitted should be identified with vet code and the tube number corresponding to
the tube number on the submission form. A mane tag is to be affixed to each horse tested and tag number recorded on
the submission form. If additional tests other than EIA are requested, please, submit two samples. The submission form
(VS form 10-11) must be completely and correctly completed and signed by the submitting veterinarian. Continued on the
next page.

Please note the following:

• KAHDL cannot make any changes or fill in any missing information.
• All forms not properly completed will be rejected and a correct form will need to be submitted.
• Samples received before 2PM will be set up for testing that day if the laboratory has sufficient staffing. Results are
normally available in 3 business days. Samples received after 2PM will be tested the following day and results will
normally be available in 4 business days from the date of drop-off.
• Submit serum tubes in slotted blood boxes. These are supplied at no cost by calling 615-837-5231. Leave indicated
information on recording.
Export testing: The submitting veterinarian is responsible for informing the laboratory of any special requirements (i.e.,
dilutions, type of test). If this information is not supplied, there may be a delay in receiving test results. For information on
test regulations, call:
• Federal – USDA, APHIS, VS, Veterinarian in charge @ 615.781.5310
• State – Office of the State Veterinarian @ 615.837.5120
When calling for results, we must have the following information exactly as provided on the submission form:
• Submitting veterinarian’s name
• Owner’s name
• Animal’s name or ID
• Sample number or ID
• Submitting date
• Bleeding date
Advance notice (7-10 days ahead) should be provided when submitting a large number of samples.
For sero-diagnosis in the individual animal, paired sera are recommended to test for specific antibody: the first sample
taken when the animal is initially examined (acute phase serum) and the second sample 2-4 weeks later (convalescent
phase serum). A rise in antibody titer between the paired samples is a basis for a specific serologic diagnosis for a particular
disease. The paired serum samples should be submitted together in order to obtain a better understanding of the
diagnostic significance of titers as they relate to the clinic status of the animal.
If only a single serum sample can be obtained, it must be taken from a convalescent animal. An acutely ill animal is
virtually devoid of antibody against the particular disease-causing agent.
If only acutely ill animals are present, swabs or tissues for isolation or culture of the causative agent should be submitted.
Submit serum tubes in slotted blood boxes. These are available at no cost by calling 615-837-5231 and leaving indicated
information on the recording.
If there are any questions regarding submission of samples, please call KAHDL at 615-837-5125.
SAMPLE SUBMISSION FAILURES
Examples of problems in submission of serum and tissue specimens are indicated below in order of decreasing frequency of
occurrence.
1. Improper sample or incomplete submission forms.

2. Serum not separated from blood clots.
3. Chemical (detergent, disinfectants, etc.) contamination of serum causing toxicity to tissue culture or otherwise
adversely affecting desired tests.
4. Hemolysis.
5. Insufficient quantity.
6. Overheating or freezing of blood before serum extraction.
7. Leaky stoppers or conatiners.
8. Broken containers or improper container.
9. Decomposed tissue specimens.

U. S. POSTAL MAILING AND PACKAGING REQUIREMENTS

FOR ETIOLOGIC BIOLOGICAL LAB SPECIMENS TO THE TENNESSEE DEPARTMENT OF AGRICULTURE
Please note that sending items by the USPS may result in significant delays – in some cases, up to a week increase in turn-
around time can be expected if the sample is still viable when USPS is used instead of FedEx® or UPS®. All etiologic and
biologic specimens sent to the Department of Agriculture through the U. S. Mail must be properly packaged so the contents
will not leak during shipment. Reference: Domestic Mail Manual, CO23.10.0-10.7.
• The specimen must be packaged in a securely sealed watertight primary container (test tube, vial, etc.) which is then
placed in a watertight, secondary plastic container with a tightly sealed closure that cannot open during shipment.
• CAUTION: Do Not Use Sealable Food Bags except for submission forms
• The specimen container and the secondary container must then be placed in a sturdy cardboard box protected with
additional absorbent packing material. The submission form should be submitted in a leak-proof sealed bag.
• Etiologic agents and Biohazard Materials must be sent by first-class Mail, Priority Mail, Express Mail, UPS® or FED-EX..
• Leak-proof Formalin containers and adsorbent materials are available from the following suppliers:
• Labsco, (800) 888-5227
• http://www.labsco.info/default.aspx?Page=Home
• Consolidated Plastics (800) 362-1000
• www.consolidatedplastics.com/Natural-And-Clear-Wide-Mouth-Threaded-Jars-C1581.aspx
• Path-tec, (706) 569-6368
• http://www.path-tec.com/products/productpages.htm
SUBCONTRACTORS
NOTE: Any test not listed in this Lab Users Guide is subject to subcontracting. Subcontracting of tests may also occur if a
test or procedure normally conducted at Kord ADDL is not currently available. Submission of samples to Kord ADDL implies
authorization to utilize testing by these laboratories. Any fee-based testing requires client preapproval. All toxicology
samples are subcontracted to another division within the Tennessee Department of Agriculture.
Subcontractor Address Phone Tests Requested Certification

NVSL 1800 Dayton Ave, USDA, 515-663-7212 Non-KAHDL ISO 17025
APHIS, Ames, IA 50010- diagnostic testing
9674
Plum Island Animal P.O. Box 848 631-323-3200 Foreign Animal ISO17025
Disease Center Greeneport, LI Orient Disease Confirmation
Point, NY 11944
University of 2407 River Drive 865-974-5673 Formalin-fixed tissue AAHA
Tennessee Dept. of Room A201
Pathobiology Knoxville, TN
37996-4542
Tennessee DOH 630 Hart LN 615-262-6351 Rabies, Salmonella External proficiency testing by
Nashville, TN typing Wisconsin Dept. of Hygiene
Toxicology Section, Ellington Agriculture Ctr, 615-837-5414 Routine toxicology Audited annually by KAHDL
Technical Services 440 Hogan Rd samples
Division, Nashville, TN 37220
Tennessee DOA
Dr. Phil Elzer, LSU, 111 Dalrymple Bldg 225-578-4763 Swine Brucellosis None, at direction of
Veterinary Baton Rouge, LA 70803 Tennessee AVIC
Immunology
Georgia Poultry 4457 Oakwood Road 770-535-5996 Poultry infectious None
Laboratory Oakwood, GA 30566 diseases
Network

ABORTIONS
Diagnosis of the cause of abortion is often difficult and complex. Tips to increase your diagnostic yield include:
• Submit tissues from multiple fetuses and placentas to increase the probability of diagnosis.
• For serum titers, collect and submit the first of paired serum samples from the suspected aborting animal and follow in
2-3 weeks with the second sample.
• If a toxic condition is suspected, submit samples of the aborting animal’s feed and water along with aqueous humor or
an intact eye.
• If the fetus and placenta cannot be submitted, submit the following:
o FRESH TISSUE for bacterial culture & virology may include stomach contents, placenta, liver, lung, spleen, kidney,
& brain.
o FIXED TISSUE for histopathological examination should include placenta, Lung, liver, heart, adrenal, kidney,
thymus, and brain.
o OCULAR FLUID (best) or eye for nitrates or if hypomagnesemia is suspected.
BACTERIOLOGY SUBMISSIONS
KAHDL does not conduct any testing for food safety purposes or make any recommendations regarding safety or edibility of
items for human consumption.
• AEROBIC SPECIMENS FROM NECROPSIED ANIMALS
o Collect all specimens as aseptically as possible. A golf ball-sized portion of each organ should be collected.
o If the outside of the specimen is accidentally contaminated, wash the specimen with clean tap water.
o Refrigerate (wet ice packs) all specimens to prevent saprophytic overgrowth.
o Collect observable lesions or suspected target organs.
o For neonatal diarrhea, submit a tied off 4-5 cm segment of jejunum, ileum, and colon with the accompanying
lymph nodes for culture of pathogenic bacteria. Fecal floatation is not performed at KAHDL.
o Tissue specimens should be placed in individual leak-proof plastic bags & identified (use water-proof ink). Ideally
each tissue should be separate - at a minimum, separate gastrointestinal tract from other organs.
• MASTITIS MILK SPECIMENS
o Wash udder to remove dirt and allow drying.
o Scrub teat end with alcohol soaked cotton and let it dry.
o Samples should be collected in a sterile container immediately prior to regular milking without discarding any
streams of milk (the first milk usually contains the greatest number of the infecting micro-organisms).
Approximately 1-2 mls is sufficient.
• SWAB SUBMISSIONS - Collect samples aseptically and submit in commercial transport media.
• ANAEROBIC AND MICROAEROPHILIC SPECIMENS
o Note: Anaerobic & microaerophilic organism culture is heavily dependent on sample selection & shipment.
o Samples should be taken from a living animal or a fresh carcass.
o Specimens should be submitted in a transport media that limits or excludes air from the sample. Use a
commercial anaerobic transport media swab.
MYCOLOGY (FUNGAL CULTURE) SUBMISSIONS
• COLLECTION AND CARE OF SPECIMENS - Currently the only fungi cultured by the laboratory are dermatophytes.
Submit skin scrapings from the outer edges of a lesion and submit plucked (not cut) hairs. Skin, hair, and nails should be
shipped to the laboratory without refrigeration.
• RESULTS - Fungal isolations normally take longer than bacteria; therefore, we recommend waiting 14-21 days before
expecting a result. Feel free to call to check on culture status if there is a question.
• DIMORPHIC FUNGI – Blastomyces and Histoplasma are among the highly pathogenic dimorphic fungi that pose a
significant risk to laboratory personnel. These diseases are best diagnosed by serological methods, cytology and/or
histopathology which provide a more rapid diagnosis than fungal culture.

TURN-AROUND TIME
• Please note that we make every effort to assure prompt and accurate results with the resources we have available.
• Some tests such as fungal and Mycobacterial culture may take several weeks or more; if there is a question
regarding results, feel free to call and we can give you an estimated finalization date.
• Examples of some of the prolonged tests/assays that require extended periods of time to finalize are:
o Mycoplasma, 7-10 days
o Dermatophytes/fungal, 14-21 days
o Johne’s disease (paratuberculosis), 1-8 weeks – reported earlier if there is a positive test result
USER’S GUIDE TO MICROBIOLOGY
The Microbiology section of the Animal Health Diagnostic Laboratory provides culture & testing for a wide variety of
infectious diseases. Some of the most common are listed below along with the preferred sample.
TEST SAMPLE
Acid Fast Stain Feces or intestine (for Mycobacterium paratuberculosis and Cryptosporidia)
Aerobic Culture Fresh chilled tissue, urine, exudate, transtracheal wash
Anaerobic Culture Fresh tissue, anaerobic culturettes, exudate
Antibiotic Sensitivity Performed on isolates recovered from specimens
(disc diffusion, Vitek®) Please request on accession form if you would like this test done
Blood Culture Blood submitted in blood culture bottle
Brucella abortus culture Fetal tissues, placenta, milk, lymph nodes
Brucella canis Fetus, testicle, placenta, uncoagulated blood in a blood culture bottle, lymph nodes, vaginal
discharge, milk, semen
Calf Scours Feces or affected intestine
Campylobacteriosis Affected intestine – must specifically request (requires specific media)
Candida Lesion, milk
Dermatomycosis Lesion, hair, scales, fungal slants/trays
Dermatophilosis Hair and scabs (please submit a generous sample)
Diarrhea/enteritis Feces, affected intestine
Johne’s Dz (M Feces (2 gram sample, Walnut-sized sample)
paratuberculosis)
Lumpy Jaw (Actinomyces) Exudate, lesion, sulfur granules
Mastitis (milk) culture Milk submitted in Whirl-Paks® or sterile tubes. Approximately 2-4 ml is sufficient.
Please NOTIFY the laboratory in advance if submitting more than 40 samples at a time
Mycoplasma Fresh chilled tissue, transtracheal wash, swab (may require 10-14 days for completion).
Paratuberculosis (Johne’s). Feces (2 gram sample, Walnut-sized sample)
Pinkeye (Moraxella bovis) Culturette of affected eye. Sometimes difficult to isolate from normal flora
Pneumonia Lung (Please indicate if Mycoplasma, Hemophilus, or Rhodococcus is suspected)
Salmonella feces, feed, water, environmental samples
Strep. equi equi (Strangles) Exudate from non-draining lesion
Trichomonas culture Preputial wash, vaginal fluid, CALL FOR PROTOCOL before submitting samples

PROTOCOL FOR TRICHOMONAS COLLECTION & SUBMISSION IN CATTLE
Note that the preferred method for diagnosing trichomoniasis in cattle is now PCR; however, some regulatory agencies may
still require culture. Diagnosis of trichomoniasis is made when Trichomonas organisms are observed from the smegma or
preputial flush samples of bulls, or the uterine/vaginal fluid from cows. All samples must be submitted in an InPouch™ TF
pouch. These pouches may be purchased from Biomed Diagnostisics.¹ Accurate diagnosis is dependent on the efficacy of
sample collection, handling, and processing. When a positive sample is found, at the client’s request, it will be submitted for
Polymerase Chain Reaction (PCR) to distinguish between T. foetus and other Trichomonas organisms. There will be an
additional charge for the PCR.
• MATERIALS REQUIRED:
o InPouch™TF pouches
o Disposable gloves
o Infusion pipette
o 20 ml syringes or wooden applicator sticks/sterile cotton-tipped swabs
• SAMPLES:
o Smegma or preputial flush of bulls
o Uterine/vaginal fluid from cows
• SAMPLE COLLECTION:
o To view online training course Trichomoniasis Testing Course for Bovine Practitioners, go to:
 http://extension.wsu.edu/vetextension/Beef/trich/Pages/default.aspx
 Contact Dr. Clariday, Assistant State Veterinarian, at 615. 837.5120 for questions about certificates of completion
for this training course.
• INOCULATION OF InPouch™: For details, see the InPouch™TF manufacturer’s instructions.
• SUBMISSION and SHIPPING REQUIREMENTS:
o Samples must be collected into InPouch™ TF pouches (see vendor info below)
o o
o Pouches must be kept at ambient (room) temperature (65-80 F or 18-27 C) prior to shipping to the lab
o Avoid overheating or freezing the samples
o DO NOT use expired InPouch™ TF pouches (dated on side of each pouch). Such samples will be rejected from testing.
o DO NOT submit samples in other containers or media or they will be rejected from testing.
o DO NOT refrigerate or freeze the inoculated InPouch™ samples, or the un-used InPouch™.
o Send the inoculated InPouch™ samples in an INSULATED CONTAINER with NO ICE PACKS by overnight express/one
day delivery (Not USPS). The Kord Laboratory should receive the samples within 24-48 hours after collection.
o Samples in transit for more than forty-eight (48) hours after collection will NOT be accepted for testing.
o When planning to collect ten (10) or more samples, please schedule the submission with the laboratory prior to
collecting the samples. Phone: 615.837.5125.
o Provide a master list of the samples with the submission by numbering the InPouch™ with consecutive numbers (1, 2,
3, etc.). Keep a master list of corresponding animal ID or tattoo numbers and send a copy to the lab along with the
regular lab submission form.
o Schedule shipments to avoid weekend or holiday delivery to the laboratory. (Samples are accepted on Friday if they
are delivered in person by 4:00 P.M.)
• TURNAROUND TIME:
o Culture: Six (6) business days after samples are received by the lab.
o PCR: Two (2) to five (5) business days after samples are received by the lab.
¹BioMed Diagnostics, Inc., 1388 Antelope Road, PO Box 2366, White City, Oregon 97503 - (800) 964-6466
• www.biomeddiagnostics.com
• InPouch™TF Test – Bovine Cat # 11-1003 100 tests
• InPouch™TF Test – Bovine Cat # 11-1010 10 tests

NECROPSY (AUTOPSY) SUBMISSIONS & CARE OF REMAINS
NOTE: Animals MUST be euthanized before submission to the laboratory.

• Cool dead animals as soon as possible after death.
o Large animals should be thoroughly hosed down with cold water.
o Birds, rabbits, & fur-bearing animals:
 soak in cold, soapy water
 place in a plastic bag
 refrigerate
o NOTE: Do not place animals in a plastic bag without prior cooling.
• A cursory confirmatory examination or refusal of an animal for necropsy at any time will be at the discretion of the
diagnostician assigned to the case and will be based upon the following:
o An animal is deemed too decomposed for further diagnostic testing
o An animal has signs that are consistent with a recent laboratory diagnosis, therefore necropsy is unwarranted
o An animal has already had a diagnosis confirmed by a veterinarian or owner and therefore a necropsy is
unwarranted. (e.g. fractured leg, uterine prolapse, or chronic laminitis)
• In all cases, the KAHDL diagnostician will determine the suitability of submitted animals, tissues, or other materials for
testing and determine which tests are performed.
• When multiple animals are submitted representing a herd or flock problem, the KAHDL diagnostician will determine the
number of animals to be necropsied.
• Legal/cruelty cases - Animal cruelty/legal cases will be handled in the same manner as a routine necropsy submission.
o Additional veterinary forensic testing such as determining the time of death, forensic entomology, or
determining types of accelerants used in burn cases will not be performed.
o Please notify the laboratory at the time of submission that the case may involve cruelty or a legal issue.
o Any pictures of the animal must be taken prior to submission of the animal to the diagnostic laboratory, as
pictures will not be taken during the gross necropsy.
• When bullet retrieval or confirmation is desired in suspected gunshot cases, radiographs must be taken and brought to
the laboratory upon submission of the animal for necropsy.
• KAHDL will not conduct any testing for food safety purposes or make any recommendations regarding safety or edibility
of items for human consumption.
• Care of remains and cremation services – KAHDL does not perform private cremations. All remains are incinerated
without return of individual animal ashes. If return of ashes is elected, they will be representative ashes from the day’s
incineration and an additional fee will be charged. If a cremation service is elected, remains are saved and packaged for
transfer to one of the local cremation services. I such cases, there is a fee for saving and handling remains for the
cremation service (in addition to their normal fees).
HISTOPATHOLOGY - COLLECTION AND CARE OF SPECIMENS
1. Diagnostic accuracy is directly proportional to the collector’s ability to select the specimen that represents the lesion or
disease process. Poor selection can result in inaccurate interpretation. Improper samples or samples deemed non-
diagnostic will not be processed. Specimens should include grossly observable lesions with a small amount of adjacent
normal tissue.
2. Specimens should be from the organ containing the lesion. Tissue specimens should include the surface and all
anatomical features; for example, specimens of the kidney must include cortex, medulla, and pelvis.
3. The entire brain should be removed and cut longitudinally on the midline into two equal portions; 1/2 should be
submitted in 10% BNF for histopathology and 1/2 submitted fresh for other test procedures, as indicated. (Gross
examination by an experienced pathologist is often necessary to locate focal lesions for further sectioning - random
samples often miss important lesions.) – THE EXCEPTION TO THIS PROCEDURE IS IN CASES WHERE RABIES IS A
DIFFERNTIAL – IN SUCH CASES, BRAIN SHOULD EITHER BE SUBMITTED DIRECTLY TO THE HEALTH DEPARTMENT OR
THE ENTIRE BRAIN SHOULD BE SUBMITTED FRESH ON ICE.

4. Specimens (except the brain) should be 0.5 cm to 1 cm thick. Specimens that are too thin cannot be properly trimmed
for sectioning and those that are too thick decompose before they are fixed. (Formalin penetrates approximately 3 mm
on each side of the section per day at room temperature.)
5. Fixation must begin as soon as possible after a carcass is opened or a surgical specimen is procured. At least ten (10)
parts of fixative to one (1) part of tissue is necessary. Samples may be shipped in a smaller volume after being fixed.
6. Buffered, formalin-fixed tissues are required for histopathology examination. Neutral-buffered formalin is available
commercially or the following recipe can be used:
• 900 cc of water
• 100 cc 40% formaldehyde
• 5 grams calcium carbonate
7. Intestinal specimens requiring critical examination of villi (i.e. rotavirus and coronavirus infection) require special
handling. The preferred method is to tie off approximately 3 cm long segments of bowel and gently fill the segments
with 10% neutral buffered formalin, using a needle and hypodermic syringe.
8. Skin, gastrointestinal endoscopic biopsies and uterine biopsy specimens should be placed on a piece of tongue
depressor, sliced cucumber (GI biopsies), or smooth cardboard
• Do not use paper
• Subcutis/submucosa or cut surface should be in contact with the wood or cardboard
9. In order to provide meaningful information for tumor margin evaluation, margins should either be inked with a
commercial inking system or should have sutures placed and a clear indication of specimen orientation should be
included with submission forms.
10. The mouth of specimen containers should be wide enough to allow the tissue to drop into the bottle without touching
the sides of the opening. Note that unfixed tissue can be easily forced into a jar that has an opening too narrow to
allow removal following fixation without breaking the container. Glass container should be avoided.
SHIPMENT
1. Use wide mouth plastic or non-breakable bottles or vials with leak-proof lids. Avoid taping containers shut; it does not
prevent leakage (Parafilm™ is acceptable). There are recycled containers available at KAHDL.
2. Refer to attached postal guidelines. Note that using the USPS instead of FedEx® or UPS® may significantly slow turn-
around times. Unless there are special requirements (decalcification or special stains), we will do everything possible to
limit turnaround time to one day to process slides and another day for them to be read by pathologists (total turn-
around from receipt of fixed tissue should be less than 2-3 days).
3. Pack the specimens with adequate padding to prevent breakage.
4. Avoid cramming large quantities of tissue into a small container.
5. Submit tissue in 10:1 ratio of fixative to tissue or fix the tissue and then transfer it to a smaller container with less
formalin for shipment.
6. For multiple samples from a single patient, samples should be clearly differentiated in separate containers or somehow
marked (ink, suture) if submitted in the same container.
SUBMISSION FORM
1. Provide ALL the requested information on the form.

2. Brief, concise, complete histories are required and aid in providing diagnoses and pertinent advice.
3. Please use black ink and write or print legibly using indelible ink (no felt-tip markers).
4. List the tissues submitted, also the number of samples. This will help insure that all submitted specimens are identified
and examined. Always indicate the source or location from which the specimen was obtained.
CYTOLOGY
• The laboratory offers cytologic and peripheral blood smear examinations. However, we do not do clinical chemistries,
CBC’s or differential blood counts. These can best be accomplished by commercial or hospital laboratories.

• Cytologic aspirates are safe, easy, and often valuable. However, cytology does have its limitations. Material collected
may not always represent the ongoing process. For example, large quantities of blood in an aspirate may represent
part of the pathologic process or be due to the aspiration procedure. Insufficient cellular material in an aspirate may
result when working with fibrous tissue such as fibrosarcomas. The quality of the sample strongly influences the
diagnostic potential of cytology. Therefore, close attention must be made to slide preparation and handling. Ideally,
preparations should be thin enough to visualize individual cells but cellularity must be sufficient for diagnosis. Cells
should be handled gently to prevent destruction. Because fresh cells make the best preparation, slides should be
prepared promptly after collection.
• Fine Needle Aspiration
1. Use a 22 gauge needle with 10-12 cc syringe and pre-cleaned slides.
2. Take several vigorous aspirates from mass.
3. In order to avoid rupturing of cells, release suction pressure before removing the needle from mass. Often the
specimen will be contained only in the hub of the needle.
4. After withdrawing the needle from the mass, remove the needle from the syringe. Then, fill the syringe with air,
replace needle and use aspirated air to force cellular material onto slide.
5. Make a “squash” or “pull-apart” smear by covering the material on the slide with another slide, squashing the
material on the slide with digital pressure and then pulling the slides apart. This must be done quickly as
cytologic material often clots rapidly. Note that excessive pressure will destroy labile cells such as neoplastic
lymphocytes.
6. Please sent 3-4 unstained, air dried smears.
Note: Lymph node aspirates must be handled gently. Lymphocytes are frequently damaged if shear force is applied
to them. This is especially true in the case of malignant lymphoblasts. Slides should be squashed together
by digital pressure and pulled apart vertically rather than horizontally to avoid shear force.
• Imprints - Imprints or touch-preps can be made from solid tissue. A fresh surface should be blotted to remove the
majority of surface blood. Several imprints per slide should be made. Material should not be smeared.
• Scrapings – Fibrous tissues are best sampled by scraping. A fresh surface is cut and then scraped using a clean scalpel
or razor blade. The material is then gently spread across the slide.
• Body fluids and washes - Slides from turbid fluid samples can be made in the same manner as peripheral blood slides.
Clear or slightly turbid fluids should be centrifuged and the sediment spread on slides. Smears must be made promptly
after each collection. Cellular degeneration will be evident within 2 - 3 hours after collection.
• Evaluation for blood parasites - Submit 2 unstained, air dried blood smears (fresh blood preferable to avoid artifacts
caused by EDTA).
• Only slides prepared at the time of collection will be examined. Do not submit only fluids or blood for microscopic
evaluation.
• Additional cytology tips can be found in our newsletter, KAHDL NOTES volume 1, number 2.
Collection procedures can also be reviewed in:

1. Baker, R & Lumsden, JH (2000). Color atlas of cytology of the dog and cat. Mosby, St Louis, Missouri.
2. Cowell, RL, Tyler, RD, Meinkoth, JH, & DeNicola, DB (2007). Diagnostic cytology and hematology of the dog and cat.
Mosby, St Louis, Missouri.
3. Perman, et al (1979). Cytology of the dog and cat. American Animal Hospital Association, South Bend, Indiana.
4. Rebar, A.H. (1978). Collection Techniques in Veterinary Cytology. In: Handbook of Veterinary Cytology. Ralston Purina
Company, St. Louis, Missouri.
PARASITOLOGY
The KAHDL no longer accepts fecal samples for fecal flotation testing. Identification of ectoparasites is available.
Cryptosporidia may be identified in a fecal smear submitted to the Bacteriology Section.

SEROLOGY & MOLECULAR DIAGNOSTICS
KAHDL personnel are certified by NVSL to perform Brucella abortus card testing, BAPA, and the standard plate test.
Brucella abortus antigen is also used for porcine testing (B suis cross reacts), as is the BAPA test (which is also used for
testing cervidae). Serology personnel are also certified to perform avian influenza AGID testing. The expected
turnaround time for selected analyses based on batch sample testing follow:
• 24 Hour turnaround is expected on agglutination tests, AGID tests, and fluorescent antibody testing
• 48 hour turnaround is expected on Immunofluorescent antibody testing
• 4 day turnaround is expected in ELISA tests and Leptospirosis microagglutination testing
• 1 week turnaround is expected on serum neutralization tests
Note that turnaround times are based on when we receive the sample. Shipping by overnight UPS® or FedEx® is always
recommended to speed up the testing process. We do not recommend USPS shipping.
Key to abbreviations for Serology Test Chart:

AGID - Agar gel Immunodiffusion CA - Card Agglutination ELISA - Enzyme Linked Immunosorbent Assay
FA - Fluorescent Antibody (Direct) HI - Hemagglutination Inhibition IFA - Indirect Fluorescent Antibody
MA – Microagglutination PA – Plate Agglutination PCR - Polymerase Chain Reaction
RSA – Rapid Serum Agglutination
SEROLOGY & MOLECULAR DIAGNOSTIC TESTS BY SPECIES (SUBMIT ALL SERUM IN BLOOD BOXES)
AVIAN (POULTRY):
TEST DAYS
TEST SPECIMEN SHIPPING TYPE RUN COMMENTS
Avian Influenza Serum (at least 1 ml) Refrigerate ELISA M-F
AGID M-Th Not run on Friday (24 hour test)
Cloacal, tracheal, &/or PCR Requires special request
oropharyngeal swab-
inoculated broth
Chlamydophyla Spleen, liver, lung, air sac, Refrigerate FA M-F Reported as positive or negative
conjunctival swab
Mycoplasma Serum Refrigerate ELISA M-F Requires at least 1 ml of serum
galliseptica & M. Tracheal swab (call first for HI Submit in appropriate transport media
synoviae submission directions) PA
Salmonella Serum (at least 1 ml) Refrigerate RSA M-F Screening test – MA used for
pullorum confirmation
MA Confirmatory test for positive RSA
results

CANINE (DOG):
TEST DAYS
Blastomycosis Serum Refrigerate AGID M-F
Brucella canis Serum Refrigerate IFA M-F Samples run at 1:10, 1:50, 1:250, 1:1250
dilutions.
Distemper Virus Lung, kidney, spleen, urinary Refrigerate FA M-F Reported as positive or negative
bladder, brain, stomach, liver,
blood smear
Serum IFA M-F IgG, IgM determination. Run at 1:10,
1:50, 1:250, & 1:1250 dilutions
Ehrlichia canis Serum Refrigerate IFA M-F Run at 1:10, 1:50, 1:250, & 1:1250
dilutions.
Herpesvirus Lung, liver, kidney, spleen, Refrigerate FA M-F Reported as positive or negative
lymph node
Serum IFA IgG, IgM determination. Run at 1:10,
1:50, 1:250 & 1:1250 dilutions.
Histoplasmosis Serum Refrigerate AGID M-F Reported as positive or negative
Hepatitis Serum Refrigerate IFA M-F IgG, IgM determination
(adenovirus) Run at 1:10, 1:50, 1:250, & 1:1250
dilutions.
Lyme Disease Serum Refrigerate IFA M-F Samples run at 1:64, 1:128, 1:256, 1:512
(borreliosis) dilutions
Leptospirosis Serum Refrigerate MA Batch Serum tested for 5 serovars - canicola,
Kidney, lung FA M-F grippotyphosa, hardjo,
icterohemorrhagiae, and pomona, at
1:100, 1:200, 1:400, 1:800, 1:1600,
1:3200, and 1:6400 dilutions. Usual
turnaround time is 4 days.
Neospora Brain, lung, liver, kidney, Refrigerate FA M-F Reported as positive or negative
caninum muscle, skin
Serum IFA Sample tested at 1:50
Parvovirus Intestine (jejunum, ileum), Refrigerate FA M-F Reported as positive or negative
spleen, mesenteric lymph
node
Serum IFA IgG, IgM determination. Run at 1:10,
1:50, 1:250, & 1:1250 dilutions
Rocky Mountain Serum Refrigerate IFA M-F Samples run at 1:64, 1:128, 1:256, 1:512
Spotted Fever dilutions

EQUINE (HORSES)
DAYS
TEST SPECIMEN SHIPPING TEST RUN COMMENTS
Brucella abortus Serum Refrigerate Card, M-F Reported as positive or negative
BAPA,
std
plate
Equine Infectious Serum Refrigerate AGID M-F Complete EIA form (VS Form 10-11).
Anemia (EIA) Submitting veterinarian’s signature is
required. Samples received before 2PM
will be set up for testing that day. The
test takes 24 hours. Submit tubes in
slotted blood boxes (Available at no
cost; 615-837-5231)
Equine Rhino- Lung, liver, spleen, fetal Refrigerate FA M-F Reported as positive or negative
pneumonitis Virus tissues
(ERV) Serum SN M,Th,F Titers begin at 1:4, 1:8, 1:16, 1:32, 1:64,
1:128: 1:256
Potomac Horse Serum Refrigerate IFA M-F Samples run at, 1:80 and 1:160 dilutions.
Fever
grippotyphosa, hardjo,
icterohemorrhagiae, and pomona, at
1:100, 1:200, 1:400, 1:800, 1:1600,
1:3200, and 1:6400 dilutions. Usual
Kidney, lung FA M-F
West Nile Virus Send to Breathitt Veterinary
Center
https://breathitt.murraystate.
edu/)

FELINE (CATS):
TEST DAYS
Chlamydiophyla Conjunctival smear, nasal Refrigerate FA M-F Reported as positive or negative
swab, lung
Feline Calicivirus Serum Refrigerate IFA M-F Samples are tested for IgG titers at 1:10,
1:50, 1:250 and 1:1250 dilutions
Feline Herpes Nasal swab, conjunctival Refrigerate FA M-F Reported as positive or negative
virus swab, tonsil, trachea, lung
Serum IFA Samples are tested for IgG titers at 1:10,
1:50, 1:250 and 1:1250
Feline Infectious Affected tissues (kidney, liver, Refrigerate FA M-F Reported as positive or negative
Peritonitis (FIP) spleen, lymph nodes)
Serum, pleural or peritoneal IFA M-F Sample is tested at 1:6400 dilution
fluid
Note: PCR testing for FIP virus in circulating monocytes is currently the test of choice for diagnosing FIP. This
testing is offered through Auburn University – see:
http://www.vetmed.auburn.edu/feline_infectious_peritonitis_virus2
Panleukopenia Small intestine, lymph node, Refrigerate FA M-F Reported as positive or negative
spleen, aborted fetus
Serum IFA Samples tested for IgG titers at 1:10,
1:50, 1:250, 1:1250 dilutions.
Toxoplasmosis Serum Refrigerate IFA M-F Samples are tested at 1:16, 1:32, & 1:64,
& 1:128 dilutions.

PORCINE (PIGS)
TEST DAYS
Kidney, lung FA M-F grippotyphosa, hardjo, icterohemorrhagiae,
and pomona, at 1:100, 1:200, 1:400, 1:800,
1:1600, 1:3200, and 1:6400 dilutions. Usual
Brucella suis Serum Refrigerate CARD Batch
BAPA M-F BAPA = Buffered Acid Plate Agglutination
Circovirus Lung, spleen, liver, kidney, Refrigerate FA M-F Results reported as positive or negative
lymph node
Serum IFA Samples are tested at 1:16, 1:32, 1:64, &
1:128 dilutions.
Porcine Fetal tissues Refrigerate FA M-F Reported as positive or negative
Parvovirus Fetal serum or fetal fluid IFA Samples run at 1:10, 1:50, 1:250 and 1:1250
dilutions. A single serum sample from the
dam is of little value since the breeding herd
is often seropositive.
Porcine Serum Refrigerate IFA M-F Sample is tested at 1:20 dilution.
Respiratory and
Reproductive
Syndrome (PRRS)
Pseudorabies Brain, Lung, Tonsil, Kidney Refrigerate FA M-F Reported as positive or negative
Serum ELISA Batch
Rotavirus Feces, intestine Refrigerate ELISA Batch Reported as positive or negative

RUMINANTS (CATTLE, SHEEP, AND GOATS)
TEST DAYS
Anaplasmosis Serum Refrigerated CA M-F Reported as positive or negative.
or Frozen** **For the card test, serum should be
refrigerated if analyzed in 48 hrs or frozen if
longer interval between collection and
testing.
Refrigerate ELISA BATCH For export, advance notice (> 1 weeK)
required
Bluetongue Serum Refrigerate AGID M-F Reported as positive or negative
Bovine Leukosis Serum Refrigerate AGID M-F Reported as positive or negative
Bovine Lung, bronchial lymph node Refrigerate FA M-F Reported as positive or negative
Respiratory
Syncytial Virus Serum Refrigerate IFA Samples run at 1:50 dilution.
TM
Bovine Preputial wash Avoid PCR BATCH Sample must be submitted in InPouch TF
Trichomoniasis temperature pouches. Results are reported as positive or
extremes negative.
Bovine Viral Lung, intestine, turbinate, Refrigerate FA M-F Reported as positive or negative
Diarrhea (BVD) trachea, lesion swabs, fetal
organs
Serum SN BATCH Titers: 1:4, 1:8, 1:16, 1:32, 1:64, 1:128, 1:256
Brucella abortus Serum Refrigerate Card M-F Reported as positive or negative – also
test applicable to camelids & other ungulates
BVD Persistent Serum, ear notch Refrigerate ELISA BATCH Follow instructions for ear notch submission.
Infection Reported as positive or negative. Analyzed
in batches as dictated by demand.
Caprine Arthritis- Serum Refrigerate AGID M-F Reported as positive or negative.
Encephalitis
Chlamydia Lymph node, aborted fetal Refrigerate FA M-F Reported as positive or negative.
tissue, joint fluid
Clostridium Gangrenous muscle, liver, Refrigerate FA M-F Test for Clostridium chauvoei and novyi.
lung Reported as positive or negative.
Epizootic Serum Refrigerate AGID M-F Reported as positive or negative
Hemorrhagic
Disease (EHD)
Infectious Bovine Lung, trachea, turbinate, Refrigerate FA M-F Reported as positive or negative
Rhinotracheitis abortus
Serum SN BATCH Titers: 1:4, 1:8, 1:16, 1:32, 1:64, 1:128,
>1:256
Johne’s Dz - Serum, bacterial culture Refrigerate ELISA BATCH Results are reported as positive or negative.
Mycobacterium PCR PCR is used as confirmatory assay for the
paratb growth on culture.
May take up to a month for PCR turn-
around.
Continued on next page

Continuation …
Leptospirosis Kidney, lung Refrigerate FA M-F
Serum MA BATCH Serum tested for 5 serovars - canicola,
grippotyphosa, hardjo, icterohemorrhagiae,
and pomona, at 1:100, 1:200, 1:400, 1:800,
1:1600, 1:3200, and 1:6400 dilutions. Usual
turnaround time is 4 days. No herd testing
— sick animals only.
Urine, kidney PCR Under development
Listeria Serum Refrigerate CA M-F Test for Type 1 and Type 4 serotypes.
Screened at 1:20.
Titers: 1:20, 1:40, 1:80, 1:160
Neospora Brain, lung, kidney, liver Refrigerate FA M-F Reported as positive or negative
caninum Serum IFA Serums run at 1:200 dilution.
Ovine Progressive Serum Refrigerate AGID M-F Reported as positive or negative
Pneumonia
Q Fever (Coxiella) Serum Refrigerate IFA M-F Samples are tested at 1:16, 1:32, 1:64, &
1:128 dilutions.
Rotavirus Feces, intestine (spiral colon, Refrigerate ELISA BATCH Reported as positive or negative
ileum, & jejunum)
Toxoplasmosis Serum Refrigerate IFA M-F Samples are tested at 1:16, 1:32, & 1:64, &
1:128 dilutions.
TOXICOLOGY
For suspected poisoning cases, samples will be subcontracted for toxicology testing. For routine toxicology testing, samples are
submitted to the Toxicology Section, Technical Services Division, Tennessee Department of Agriculture. Submission to Kord ADDL
implies authorization for subcontracting to this laboratory. The client is responsible for any costs associated with toxicology testing.
For samples sent to another laboratory for toxicology testing at the client’s request, the client is responsible for the cost. When
poisoning is suspected, please notify the laboratory at the time of submission. Please specify on submission forms which test
should be run on which tissue – just indicating “toxicology” is not sufficient (for example, “test liver for anticoagulant
rodenticides”). When submitting serum, the serum should be removed from the blood clot or a serum separator tube should be
used.
TOXICOLOGY TEST OFFERINGS
Test Minimum Sample Required Comments

Aflatoxin 20 grams feed (grain) No hay or silage - corn
Anticoagulants 5 ml serum, blood, plasma Include twelve common anticoagulant rodenticides
10 g bait is good Avoid submitting samples in medicine bottles
10 g stomach contents Do not freeze sample
10 g liver (best)
Arsenic 5 g liver and kidney Liver or kidney is the preferred sample from a dead animal
5 ml urine If from live animal (best):
5 ml blood Feed
5 g stomach contents Blood (if very high levels)
5 ml water Stomach contents (if recent ingestion)
5 g feed


BUN 2 ml ocular fluid
2 ml serum or plasma
Calcium 2 ml ocular fluid
2 ml serum
Carbamate 5 g stomach contents Avoid submitting samples in medicine bottles. Avoid plastic containers.
(Pesticide 5 g rumen contents Glass is preferred.
Screen) 5 g bait
10 g feed
Copper 5 ml serum Liver is preferred over kidney.
5 g liver or kidney
20 g feed
Cyanide Plants with cyanogenetic Samples should be quick frozen as soon as possible for shipment to the
potential lab.
i.e. sorghums, Sudan grass, corn
1 lb. dry plants or 5 lb. wet plants
10 ml blood
50 g muscle (heart) - best
Rumen contents - best
Dicumarol Feeds containing sweet clover
5 lb. wet plants or 1 lb. dry plants
Drug Screen 10 - 20 ml serum, whole blood Avoid submitting samples in medicine bottles.
(plasma) Urine is the preferred sample. *See end, for list of drugs included in
10 - 20 ml urine - best screen.
10 g bait, pills or 10 g stomach
contents
Ethylene glycol 3 ml urine Note that ethylene glycol is absorbed from the gastrointestinal tract within
3 ml serum, plasma seconds to minutes – stomach contents will only be positive for a couple
5 g stomach contents minutes after exposure and usually are of little use when clinically ill or at
5 g bait post-mortem.
Fumonisin 20 g feed No hay or silage - corn
Gossypol, free 1 lb. feed containing cottonseed
Iron 10 ml serum No hemolyzed samples!
Lead 5 ml blood, (EDTA, heparin) Heparin is preferred.
5 g liver and kidney Submit both liver and kidney.
5 ml water Kidney is better than liver.
5 g stomach contents
Magnesium 2 ml ocular fluid or 2 ml serum
Nitrate 2 ml ocular fluid
(qualitative)
Nitrate 1 lb. dry forage or 5 lb. wet
(quantitative) forage
1 pt. water
Ochratoxin 20 g feed (grain) No hay or silage.
Organochlorines 5 g stomach contents, rumen Avoid submitting samples in medicine bottles. Avoid plastic containers.
(pesticide contents, feed, or bait Glass is preferred.
screen)
Organophosphat 5 g stomach contents, rumen Avoid submitting samples in medicine bottles. Avoid plastic containers.
es contents, feed, or bait Glass is preferred.
(pesticide
screen)


Potassium 2 ml ocular fluid or 2 ml serum or
2 ml CSF
Selenium 10 g liver or kidney;10 ml whole Whole blood is preferred over serum.
blood or serum
Sodium 2 ml ocular fluid, serum, or urine
Strychnine 5 g stomach contents Avoid submitting samples in medicine bottles.
5 ml urine, serum, or blood
10 g bait
Urea 1 lb. feed
Vomitoxin 20 g feed (grain) No hay or silage.
Zearalenone 20 g feed No hay or silage – corn & small grains – hyperestrogenism.
Zinc 5 g liver, kidney, or stomach Serum sample should be in Royal blue top Vacutainer® tube for trace
contents metal analysis.
5 ml serum or 10 g feed
*DRUG SCREENS:
Drug Screen A Drug Screen B

Acetaminophen Methadone Amobarbital
Amitriptyline Methaqualone Aprobarbital
Amphetamine Morphine Barbital
Caffeine Nicotine Butabarbital
Chlorpromazine Nortiptyline Diazepam
Cocaine PCP Ethinamate
Codeine Propoxyphen Glutethimide
Diazepam Pseudoephedrin Pentobarbital
Doxepin Quinine Phenobarbital
Imipramine Strychnine Phenytoin
Meperidine Trifluopromazine Secobarbital
Meprobamate Trimeprazine
Methamphetamine
Call the lab if drug of interest is not listed.

10 grams of tissue is approximately equal in size to a golf ball.

VIRUS ISOLATION IS NO LONGER PERFORMED AT KAHDL
We recommend the Breathitt Veterinary Center in Hopkinsville, KY for virus isolation:

https://breathitt.murraystate.edu/
OTHER SOURCES OF HELP IN TENNESSEE
State Veterinarian, Dr Charles Hatcher – regulatory, interstate shipping, health certificates 615.837.5120
West Tennessee Animal Diagnostic Lab @ UT-Martin 731.881.7952
University of Tennessee College of Veterinary Medicine 423.974.8387
Necropsy, days 865.974.5673
Necropsy, after-hours 865.974.5701
Tennessee Wildlife Resources Agency (TWRA) 615.781.6500
Tennessee Department of Health: http://health.state.tn.us/contact.htm
Dr Heather Henderson, Veterinary Epidemiologist (rabies information) 615.741.3111
615.741.7247
USDA-APHIS Veterinary Services (“The Feds”) 615.781.5310
State/Federal Brucellosis Records (Brucellosis Results) 615.837.5120
Rabies Laboratories:
Use USPS specimen mail
Nashville:
Tennessee Dept of Health Lab Services, 630 Ben Allen Rd, Nashville, TN 37247
Mail: PO Box 305130, Nashville, TN 37230-5130 615.262.6350
Jackson Branch Rabies Laboratory:
Tennessee Department of Health, 295 Summar Avenue, Jackson, TN 38302-0849 901.423.6600
Knoxville Branch Rabies Laboratory (East Tennessee Regional Office):
1522 Cherokee Trail, POB 59019, Knoxville, TN 37950-9019 423.549.5201
ANIMAL POISON CONTROL CENTER - ASPCA

For any animal poison-related emergency, 24 hours a day, 365 days a year. If you think that your pet may
have ingested a potentially poisonous substance, call (888) 426-4435. A $60 consultation fee will be applied
to your credit card (they are not supported by state or national resources, but this is the best source for
animal poisonings).
TENNESSEE POISON CONTROL CENTER (HUMAN POISONINGS)

Tennessee Poison Center (TPC) provides immediate treatment advice for poison emergencies by calling the
Poison Help hotline at 1-800-222-1222. They are available 24 hours a day, 7 days a week. TPC also provides
information about poisons and poison prevention.
TPC can help you with questions about household products, chemicals at work or in the environment, drugs
(prescription, over-the-counter, herbal & illegal), snake and spider bites, & chemical terrorism. A specially
trained nurse, pharmacist or doctor will help. All calls are free and confidential.

REPORTABLE DISEASES
The reporting of evidence of certain animal diseases is a requirement under Standards for Accreditation of
Veterinarians in Tennessee and other states. Such reporting is also required by State law in Tennessee. Most
important, perhaps, is the veterinarian’s professional responsibility to report these diseases properly to ensure
that appropriate control measures may be instituted. Reportable diseases in general include all diseases for which
control or eradication programs are in effect, and all foreign diseases (not known to exist in this country).
Reportable diseases currently include but are not limited to the following:
Species Avian Equine Bovine Sheep & Goat Porcine Dog & Cat
Fowl Plague X
Fowl Typhoid X
Pullorum Dz X
Velogenic Viscerotropic Newcastle Dz X
Anthrax X X X X
Brucellosis X X
Psoroptic Scabies X
Screw Worm X X X X
Tuberculosis X
Vesicular Dzs X X X X
BSE X
Bluetongue X
Rabies X X X X X
Scrapie X
Scabies X
Equine Infectious Anemia X
Piroplasmosis X
Viral Encephalitis X
African Swine Fever X
Hog Cholera X
Pseudorabies X
Rabies is reportable in all species - Suspected or known rabies infection should also be reported to local public
health authorities. Reporting should include telephone or written notice to the State Veterinarian’s office and
submission of samples to the appropriate laboratory.





Diagnostic Laboratory Guide

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Diagnostic Laboratory Guide

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Tennessee Department of Agriculture Regulatory Services

The Kord Animal Health Diagnostic Laboratory

KORD ANIMAL HEALTH DIAGNOSTIC LABORATORY

U.S. POSTAL ADDRESS OVERNIGHT DELIVERY ADDRESS

Kord Animal Health Laboratory Kord Animal Health Laboratory

KAHDL Lab Use Guide, August, 2013 Page 1 of 28

Hours ……. ….………………………………………………………………..………………….. 2

• 8:00 AM to 4:30 PM, Central Time.

FEES (SUBJECT TO CHANGE, EFFECTIVE JANUARY 1, 2012)

DETERMINATION OF FEES CHARGED PER SAMPLE BASED ON TESTING REQUESTED/REQUIRED

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GUIDELINES FOR SAMPLE SUBMISSION

SAMPLE SUBMISSION BEST PRACTICES

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Please note the following:

SAMPLE SUBMISSION FAILURES

1. Improper sample or incomplete submission forms.

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U. S. POSTAL MAILING AND PACKAGING REQUIREMENTS

Subcontractor Address Phone Tests Requested Certification

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MYCOLOGY (FUNGAL CULTURE) SUBMISSIONS

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USER’S GUIDE TO MICROBIOLOGY

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PROTOCOL FOR TRICHOMONAS COLLECTION & SUBMISSION IN CATTLE

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NECROPSY (AUTOPSY) SUBMISSIONS & CARE OF REMAINS

NOTE: Animals MUST be euthanized before submission to the laboratory.

HISTOPATHOLOGY - COLLECTION AND CARE OF SPECIMENS

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1. Provide ALL the requested information on the form.

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Collection procedures can also be reviewed in:

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SEROLOGY & MOLECULAR DIAGNOSTICS

Key to abbreviations for Serology Test Chart:

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RUMINANTS (CATTLE, SHEEP, AND GOATS)

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TOXICOLOGY TEST OFFERINGS

Test Minimum Sample Required Comments

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Test Minimum Sample Required Comments

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Test Minimum Sample Required Comments

Drug Screen A Drug Screen B

Call the lab if drug of interest is not listed.

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VIRUS ISOLATION IS NO LONGER PERFORMED AT KAHDL

We recommend the Breathitt Veterinary Center in Hopkinsville, KY for virus isolation:

OTHER SOURCES OF HELP IN TENNESSEE

ANIMAL POISON CONTROL CENTER - ASPCA

TENNESSEE POISON CONTROL CENTER (HUMAN POISONINGS)

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